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Biomedical subjects

X Banse

Publications and source records attributed to X Banse.

15 recordsLinked to original sources

The genetic influence on bone susceptibility to fluoride.

INTRODUCTION: The influence of genetic background on bone architecture and mechanical properties is well established. Nevertheless, to date, only few animal studies explore an underlying genetic basis for extrinsic factors effect such as fluoride effect on bone metabolism. MATERIALS AND METHODS: This study assessed the effect of increasing fluoride doses (0 ppm, 25 ppm, 50 ppm, 100 ppm) on the bone properties in 3 inbred mouse strains that demonstrate different susceptibilities to developing enamel fluorosis (A/J a "susceptible" strain, 129P3/J a "resistant" strain and SWR/J an "intermediate" strain). Fluoride concentrations were determined in femora and vertebral bodies. Bone mineral density was evaluating through DEXA. Finally, three-point bend testing of femora, compression testing of vertebral bodies and femoral neck-fracture testing were performed to evaluate mechanical properties. RESULTS: Concordant with increasing fluoride dose were significant increases of fluoride concentration in femora and vertebral bodies from all 3 strains. Fluoride treatment had little effect on the bone mineral densities (BMD) in the 3 strains. Mechanical testing showed significant alterations in "bone quality" in the A/J strain, whereas moderate alterations in "bone quality" in the SWR/J strain and no effects in the 129P3/J strain were observed. CONCLUSION: The results suggest that genetic factors may contribute to the variation in bone response to fluoride exposure and that fluoride might affect bone properties without altering BMD.

Animals↗

Vertebral cancellous bone turn-over: microcallus and bridges in backscatter electron microscopy.

Backscatter electron microscopy (BSE) is a powerful technique for investigating cancellous bone structure. Its main function is to offer information regarding the degree of mineralization of the tissue within individual trabeculae. To illustrate the qualitative information that can be drawn from BSE imaging technique, we present a study on human vertebral cancellous bone. This tissue is continuously remodeled through osteoclastic resorption and osteoblastic new bone apposition. It is thought that osteoclastic resorption pits are especially deleterious for vertebral bone architecture since they often perforate the thin trabeculae; the osteoblasts being unable to repair the gap. In addition, excessive stress may also disrupt the architecture in case of trabecular fracture or damage accumulation. Waves of new bone formation were easy to identify in BSE. Often these waves were connecting both edges of a perforation and called bridges. Additionally, we present a few images of microcallus formations. A microcallus is described as a small mass of woven bone that generally repairs a trabecula. The microstructural aspects of different microcalluses are presented and discussed. Both bridges and microcallus should be considered as examples of the repair porcess since they obviously preserve the connectivity of the trabeculae. However, bridges were much more frequent than microcallus (396 vs 15). Both mechanisms probably illustrate the normal response to different local stimuli.

Adult↗

Irreversible perforations in vertebral trabeculae?

UNLABELLED: In human cancellous bone, osteoclastic perforations resulting from normal remodeling were generally considered irreversible. In human vertebral samples, examined by backscatter electron microscopy, there was clear evidence of bridging of perforation defects by new bone formation. Hence trabecular perforations may not be irreversible. INTRODUCTION: Preservation of the trabecular bone microarchitecture is essential to maintain its load-bearing capacity and prevent fractures. However, during bone remodeling, the osteoclasts may perforate the platelike trabeculae and disconnect the structure. Large perforations (>100 microm) are generally considered irreversible because there is no surface on which new bone can be laid down. In this work, we investigated the outcome of these perforations on human vertebral cancellous bone. MATERIALS AND METHODS: Using backscatter electron microscopy, we analyzed 264 vertebral bone samples from the thoracic and lumbar spine of nine subjects (44-88 years old). Nine fields (2 x 1.5 mm) were observed on each block. Several bone structural units (BSUs) were visible on a single trabecula, illustrating a dynamic, historical aspect of bone remodeling. A bridge was defined as a single and recent BSU connecting two segments of trabeculae previously separated by osteoclastic resorption. They were counted and measured (length and breadth, microm). RESULTS AND CONCLUSION: We observed 396 bridges over 2376 images. By comparison, we found only 15 microcalluses on the same material. The median length of the bridge was 165 microm (range, 29-869 microm); 86% being longer than 100 microm and 35% longer than 200 microm. Their breadth was 56 microm (range, 6-255 microm), but the thinnest were still in construction. Bridges were found in all nine subjects included in the study, suggesting that it is a common feature of normal vertebral bone remodeling. These observations support the hypothesis that perforation could be repaired by new bone formation, and hence, might not be systematically irreversible.

Adult↗

Patient-specific microarchitecture of vertebral cancellous bone: a peripheral quantitative computed tomographic and histological study.

This study directly compares peripheral quantitative computed tomography (pQCT) and histology for the assessment of 11 morphological parameters. Sixty-eight cylindrical cancellous bone samples were cored from the thoracic (T-9) thoracolumbar (T-12 or L-1), and lumbar (L-4) vertebral bodies of nine autopsy subjects (aged 44-88 years). Four transverse slices were acquired by pQCT from the bottom to the top of each cylinder. Slice thickness was 300 microm and pixel size was 70 x 70 microm. Thin sections (5 microm) were obtained at the same location in the samples, stained with Von Kossa, and photographed. Classical morphological parameters and strut analysis parameters were measured on all images (272 pQCT and 272 matched histological sections). Because of the partial volume effect and specific thresholding procedure, pQCT overestimated the absolute value of the bone volume fraction (BV/TV) and trabecular thickness (Tb.Th) by a factor 2. The trabecular number (Tb.N), trabecular spacing (Tb.Sp), and total strut length (TSL) were correctly estimated. However, the direct correlation between pQCT and histology was excellent (r2 > 0.85, p < 0.001) for BV/TV, Tb.N, Tb.Sp, TSL, and star surface. For Tb.Th, number of nodes, and number of free ends, the correlation was also good (r(2) > 0.6, p < 0.001). Using a random regression model, we also explored the ability of these parameters to add structural information to the readily available BV/TV or apparent density. The model identified significant (p < 0.001) differences between subjects. For a given BV/TV, some patients had more trabeculae (Tb.N) that were thinner (Tb.Th) and more disconnected (higher free ends and star). This was observed for both histology and pQCT morphometrical data. Our analysis demonstrates the capacity of both histology and pQCT to detect subjects with specific structural patterns in vertebral cancellous bone.

Adult↗

Cross-link profile of bone collagen correlates with structural organization of trabeculae.

Little is known regarding the mechanisms that govern the structural organization of cancellous bone. In this study, we compare the nature of the collagen in vertebral cancellous bone with the structural organization of its trabecular network. Cylindrical specimens of cancellous bone from vertebrae were obtained from nine autopsy subjects (ages 46-88). In each subject, eight pairs of corresponding samples were obtained from three levels in the spine and three areas within the vertebral body, leading to a total of 68 pairs of samples. The cylinders from one side were used for morphometry and the classical morphometrical parameters were obtained (BV/TV, bone volume fraction; Tb.Th, trabecular thickness; Tb.N, number; Tb.Sp, trabecular spacing) and strut analysis (TSL, total strut length; Nd, number of nodes; Fe, number of free-ends). The amount of osteoid bone was also quantified. The cylinders from the other side were powdered and used for collagen assessment, including the amount of collagen (% w/w), and its content in immature cross-links; such as hydroxylysinonorleucine (mol/mol of collagen) and dihydroxylysinornorleucine, as well as stable mature cross-links, such as hydroxylysylpyridinoline (HP), lysylpyridinoline (LP), and the pyrrole cross-links. A random regression model was used to explore the correlations. None of the biochemical parameters correlated with the BV/TV except the ratio between immature and mature cross-links (eta(2) = 0.34, p < 0.05). There was no relationship between the amount of osteoid bone and the cross-link profile. However, the concentration of pyrrole and HP cross-links in the bone samples correlated with the structural organization of its trabeculae, but in an opposite direction. Hence, the pyrrole/HP ratio was a good predictor of Tb.Th, Tb.N, Tb.Sp, and TSL (eta(2) > 0.65 and p < 0.01) as well as Fe and star marrow space (eta(2) > 0.45 and p < 0.05). The cylinders from subjects with high pyrrole or low HP in their bone collagen had a relatively thick and simple structure. Those with low pyrrole and high HP had relatively thin trabeculae that were more numerous and spread over a complex network. The relative concentrations of the pyrrole and pyridinoline cross-links appear to reflect the structural organization of the trabeculae.

Adult↗

Mechanical properties of adult vertebral cancellous bone: correlation with collagen intermolecular cross-links.

Although the mechanical strength of cancellous bone is well known to depend on its apparent density, little is known about the influence of other structural or biochemical parameters. This study specifically investigates the cross-linking of the collagen in human vertebral bone samples and its potential influence on their mechanical behavior. Multiple cylindrical samples were cored vertically in the vertebral bodies of nine subjects (aged 44-88 years). Three spinal levels (T9, T12 or L1, and L4) and three sample sites within a vertebral body (anterior, posterior, and lateral) were used, for a total of 68 samples. The density was measured with peripheral quantitative computed tomography (pQCT) and all cylinders were mechanically tested in compression. After mechanical testing, they were unmounted and used for biochemical analysis. The amount of collagen (wt/wt of bone) and its content in reduced immature cross-links, that is, hydroxylysinonorleucine (HLNL, mol/mol of collagen) and dihydroxylysinornorleucine (DHLNL), as well as stable mature cross-links, that is, hydroxylysyl-pyridinoline (HP), lysyl-pyridinoline (LP), and pyrrole cross-link were determined for each cylinder. None of the biochemical parameters correlated to the density. On multiple linear regression, the prediction of the mechanical properties was improved by combining density data with direct collagen cross-link assessment. The HP/LP ratio appeared as a significant predictor to the strength (r = 0.40; p = 0.001) and stiffness (r = 0.47; p < 0.001) samples with a high HP/LP ratio being stronger and stiffer. Additionally, the ultimate strain correlated to the HP or LP concentration (r = 0.38 or 0.49; p < 0.01). Different subjects had different HP/LP ratios and different HP or LP concentrations in their vertebral bone samples, and the location of origin within a subject had no influence on the concentration. These observations suggest that the nature of the organic matrix in adult vertebral bone is variable and that these variations influence its mechanical competence.

Adult↗

Does peripheral quantitative computed tomography ignore tissue density of cancellous bone?

The purpose of this work was to determine the capacity of peripheral quantitative computed tomography (pQCT) to accurately measure the true physical properties of vertebral cancellous bone samples and to predict their stiffness. pQCT bone mineral density (BMD) was first measured in ideal conditions. Ten cubic specimens of vertebral cancellous bone (10 x 10 x 10 mm) were washed with a water jet, defatted, and scanned in saline after elimination of air bubbles; thirteen slices were obtained. Seventy-one unprepared cylindrical samples were scanned in more realistic conditions, which allow further biomechanical testing. After extraction from the vertebral body, the samples were pushed into a plastic tube (no effort was made to remove the marrow or air bubbles), and only four slices were obtained to reduce the duration of scan. For the 81 samples, the true bone volume fraction (BV/TV, %), true apparent density (rho(app), g/cm(3)), and tissue density (rho(tiss), g/cm(3)) (an indicator of the degree of mineralization of the matrix) were then measured using Archimedes principle. rho(app) was closely correlated to BV/TV (r(2) = 0.97). rho(tiss) (1.58 +/- 0.08 g/cm(2)) was almost constant but had some influence on rho(app) (r(2) = 0.03, p < 0.001). The pQCT BMD predicted accurately rho(app) (r(2) = 0.96) and BV/TV (r(2) = 0.93) for the cylinders. For the cubes, in ideal conditions, the same correlations were even better (r(2) > 0.99, both). Analysis of covariance indicated no difference (p > 0.05) in the regressions due to preparation of the samples. The stiffness was better predicted by the true rho(app) (r(2) = 0.87) than by BV/TV (r(2) = 0.83), indicating that stiffness was influenced by small differences in the tissue density. Consequently, the correlation between pQCT BMD and stiffness was excellent (r(2) = 0.84). The fact that pQCT did not ignore this tissue density information compensated for the inaccuracies linked to realistic scanning conditions of the cylinder.

Adult↗

In vitro evaluation of acute cytotoxicity of human chemically treated allografts.

In order to minimize the risk of contamination associated with tissue transplantation, tissue banks commonly chemically treat the tissues whenever possible. As viral inactivation uses agents lethal to microorganisms, it is imperative to assure that chemically inactivated tissue remains biocompatible. In vitro assays can be an effective means to assess the acute cytotoxicity of chemically treated human allografts. We have used different types of cells cultured in the presence of treated tissue extract. A standard cell line, a human fibroblast (WI38), which was the same for all the samples, was chosen. In addition, as the banked tissues (bone and fascia lata) were prepared to be used in bone or as a dura mater substitute, two other cell types were also used: an osteoblastic cell line (SaOS-2) and a neuronal cell line (Neuro 2A). Cytotoxic assessment was performed by qualitative evaluation of cell morphology based on confluence, granulation, vacuolization and swelling analysis. In addition, quantitative methods based on the use of neutral red (NR) and 3- (4,5-dimethylthiazol-2-yl)-2,5 diphenyl tetrazolium bromide (MTT) were assayed. Qualitative and quantitative evaluation of fascia lata and bone extracts did not show deleterious effects on cell cultures. These results show that in vitro methods can be appropriate to select a non-toxic procedure before it is used in the human body and that several strong chemical treatments can result in a tissue suitable for human.

Journal Article↗

Inhomogeneity of human vertebral cancellous bone: systematic density and structure patterns inside the vertebral body.

In the spine, cancellous bone quality is usually assessed for the whole vertebral body in a transverse central slice. Correct identification and assessment of the weakest parts of the cancellous bone may lead to better prediction of fracture risk. The density and structural parameters were systematically investigated inside the thoracic (T-9), thoracolumbar (T12-L1), and lumbar (L-4) vertebral bodies of nine subjects. On both sides of the median sagittal plane, anterior and posterior 8.2 mm vertical cores were harvested in the thoracic vertebra. In the thoracolumbar and lumbar vertebrae, external samples were also cored. Peripheral quantitative computed tomographic (pQCT) density analysis of the 136 cores was performed at four different levels, from the lower to the upper endplate. The relatively thin slice thickness (300 microm) and small pixel size (70 microm x 70 microm) was considered sufficient to investigate the structural parameters on the four transverse slices and in the sagittal and coronal planes (total of 816 images). Using a constant threshold a binary image was generated and the morphometric data were extracted. The binary image was further skeletonized and classical strut analysis was performed. Cancellous bone density was 20% higher in the posterior cores than in the anterior and external cores. Moreover, clear vertical inhomogeneity was noted because the lowest half of the vertebral body presented lower density than the upper half (differences ranging from 25% to 15%). All structural parameters were strongly dependent on the location of the measurement. Structural differences between anterior, posterior, and external areas were mild and followed the density patterns. On the other hand, vertical inhomogeneity of the structural parameters was important. For example, in the thoracolumbar and lumbar vertebrae, the numbers of nodes or node-to-node struts were almost twofold higher in the inferior half than in the superior half (p < 0.01), whereas trabecular thickness and number of free-ends presented a center/close-to-endplate structural pattern, with central trabeculae being 15% thicker (p < 0.05) and presenting 30% fewer free-ends (p < 0.01) than the close-to-endplate ones. Variability of density and structural parameters was high and a substantial part of this variability could be explained by the place inside the vertebral body where the measurement was made. The weak part was not in the center of the body but in its upper half where the lower density did not seem to be compensated by a higher structural architecture. Further clinical investigation could enhance fracture prediction by tracking and focusing on the weakest part of the vertebral body.

Adult↗

Stiffness and compactness of morselized grafts during impaction: an in vitro study with human femoral heads.

The mechanical properties of the impacted material is important when using impaction bone grafting in revision arthroplasty. We did an in vitro study to monitor the stiffness and compactness of morsellized bone grafts during impaction. Using human osteoarthrotic femoral heads as the bone source we prepared 3 types of morselized grafts with the same bone mill: (1) purely cancellous grafts, (2) cortico-cancellous grafts and (3) cortico-cancellous bone with cartilage remnants. 5 g of bone samples were progressively impacted up to 150 times in a contained cavity. All types of grafts became stiffer and stiffer during the first 30 impactions. After 30 impactions, the grafts' compactness continued to increase slowly without concomitant changes in their stiffness. Over-impaction was not useful, but did not jeopardize the implant stability. The presence of cartilage remnants considerably compromised the increase in stiffness. In contrast, the addition of cortical bone from the femoral neck did not affect the impaction and resultant stiffness.

Aged↗

Effect of freeze-drying and gamma irradiation on the mechanical properties of human cancellous bone.

Freeze-drying and gamma irradiation are commonly used for preservation and sterilization in bone banking. The cumulative effects of preparation and sterilization of cancellous graft material have not been adequately studied, despite the clinical importance of graft material in orthopaedic surgery. Taking benefit from the symmetry of the left and right femoral heads, the influence of lipid extraction followed by freeze-drying of a femoral head and a final 25-kGy gamma irradiation was determined, with the nonirradiated, nonprocessed counterpart as the control. Five hundred and fifty-six compression tests were performed (137 pairs for the first treatment and 141 pairs for the second). Mechanical tests were performed after 30 minutes of rehydration in saline solution. Freeze-dried femoral heads that had undergone lipid extraction experienced reductions of 18.9 and 20.2% in ultimate strength and stiffness, respectively. Unexpectedly, the work to failure did not decrease after this treatment. The addition of gamma irradiation resulted in a mean drop of 42.5% in ultimate strength. Stiffness of the processed bone was not modified by the final irradiation, with an insignificant drop of 24%, whereas work to failure was reduced by a mean of 71.8%. Freeze-dried bone was a bit less strong and stiff than its frozen control. Its work to failure was not reduced, due to more deformation in the nonlinear domain, and it was not brittle after 30 minutes of rehydration. Final irradiation of the freeze-dried bone weakened its mechanical resistance, namely by the loss of its capacity to absorb the energy (in a plastic way) and a subsequent greater brittleness.

Biomechanical Phenomena↗

Ethanol treatment of tendon allografts: a potential HIV inactivating procedure.

The penetration rate of ethanol in human tendons was studied to in order to define the conditions which were necessary to achieve an inactivating concentration against the Human Immunodeficiency Virus (HIV) within the tendon. The rate of alcohol penetration was found to be slow and did not differ with different types of tendons. An average ethanol concentration of 14% (v/v) was measured in human tendons after they had been immersed for 2 h in 70% (v/v) ethanol, and a concentration of 19% (v/v) was reached after 3 h. Ethanol immersion of human tendons may represent an additional safety procedure in inactivating the HIV virus provided the duration is at least 3 h.

Anti-Infective Agents, Local↗

Comparative left-right mechanical testing of cancellous bone from normal femoral heads.

In order to assess the potential influence of various physical or chemical treatments on bone grafts, and in particular, on femoral heads to be used as bone grafts, the mechanical symmetry of the femoral head was investigated. Pairs of proximal human femora were harvested and transversally sliced using a coordinate system to orient correctly the head. After being embedded and polished, bone slices were loaded to failure between two steel columns (6 mm of diameter) in a sequential and symmetrical manner for both heads. From ten pairs, 592 mechanical assays on the cancellous bone of femoral heads were performed. There was a high variation in the mechanical properties of the specimens from the different donors. However, no statistical difference could be observed within each pair when side-to-side symmetry was tested for the different mechanical characteristics. Ultimate stress at failure and stiffness were also highly correlated for each mirror location of each pair. Provided that an accurate and reproducible method of slicing femoral head can be achieved, an excellent mechanical symmetry is observed using a static compressive test. Such a method could be used to determine the mechanical influence of any treatment applied to bone.

Aged↗

Tibial tubercle elevation with bone grafts. A comparative study of autograft and allograft.

We retrospectively investigated the fate of bone auto- and allografts in 64 patients who underwent a tibial tubercle elevation with bone graft. Half of them received an autograft and the other half, an allograft that had been processed and freeze-dried. The two groups had similar preoperative characteristics concerning age, sex and pathology. Roentgenograms were reviewed by three independent observers and scored for fusion, resorption and collapse. Clinical charts were analysed for different variables. The overall radiological score for both groups did not differ statistically. Comparison of graft fixation with one or two screws demonstrated more bone resorption in the case of a single-screw fixation. In such a case, the occurrence of a preoperative tubercle fracture had a significant adverse influence, due to a less stable fixation. From the clinical charts review, only the mean stay at hospital was significantly shorter when an allograft was performed. A bone allograft appears to be suitable to maintain an osteotomy but requires a more careful surgical technique fixation to obtain a similar result to an autograft.

Adolescent↗